Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: HO Yuan Heng Trevor
Date created: 2011-09-14 11:00:00
Date modified: 2015-05-08 01:12:34
Heat sensitive origin of replication (oriR101 & repA101-ts)
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_K524000_sequence (Version 1) |
Description
OriR101 is an low copy origin of replication that is initiated by the protein repA101. repA101-ts, a mutated form of repA101 would denature at temperatures above 37??C. Hence plasmids using this construct to drive replication can only initiate replication when cultured below 37??C (recommended <30??C). Production of repA101-ts is drived by an unknown constitutive promoter included in the construct.Notes
To make this part compatible with Biobrick standard assembly protocols, overlapping PCR was used to mutate a SpeI cut site originally present inside the construct to render it nonfunctional.Source
From the plasmid pKD46 maintained inside the E.coli strain BW25113, courtesy of E.coli Genetic Resources at Yale CGSC, The Coli Genetic Stock Center.| Sequence Annotation | Location | Component / Role(s) |
| orf60a CDS repA101-ts AT Rich region_oriR101 Promoter region of repA101ts point mutation, substitute of C as G to remove SpeI site par region regions not necessary for origin function | 2,184 373,1323 1492,1574 1329,1354 1391,1391 1721,2067 1,372 | feature/cds CDS CDS feature/cds feature/dna sequence_feature feature/promoter promoter feature/mutation sequence_alteration feature/dna sequence_feature sequence_feature feature/misc |